In my relatively short career (approximately 12 years – wait, how long?) in bioinformatics, I have been involved to a greater or lesser degree in a number of standards efforts. It started in 1999 at the EBI, where I worked on the production of the protein sequence database UniProt. Now, I’m working with systems biology data and beginning to look into synthetic biology. I’ve been involved in the development (or maintenance) of a standard syntax for protein sequence data; standardized biological investigation semantics and syntax; standardized content for genomics and metagenomics information; and standardized systems biology modelling and simulation semantics.

(Bear with me – the reason for this wander through memory lane becomes apparent soon.)

How many standards have you worked on? How can there be multiple standards, and why do we insist on creating new ones? Doesn’t the definition of a standard mean that we would only need one? Not exactly. Take the field of systems biology as an example. Some people are interested in describing a mathematical model, but have no need for storing either the details of how to simulate that model or the results of multiple simulation runs. These are logically separate activities, yet they fall within a single community (systems biology) and are broadly connected. A model is used in a simulation, which then produces results. So, when building a standard, you end up with the same separation: have one standard for the modelling, another for describing a simulation, and a third for structuring the results of a simulation. All that information does not need to be stored in a single location all the time. The separation becomes even more clear when you move across fields.

But this isn’t completely clear cut. Some types of information overlap within standards of a single domain and even among domains, and this is where it gets interesting. Not only do you need a single community talking to each other about standard ways of doing things, but you also need cross-community participation. Such efforts result in even more high-level standards which many different communities can utilize. This is where work such as OBI and FuGE sit: with such standards, you can describe virtually any experiment. The interconnectedness of standards is a whole job (or jobs) in itself – just look at the BioSharing and MIBBI projects. And sometimes standards that seem (at least mostly) orthogonal do share a common ground. Just today, Oliver Ruebenacker posted some thoughts on the biopax-discuss mailing list where he suggests that at least some of BioPAX and SBML share a common ground and might be usefully “COMBINE“d more formally (yes, I’d like to go to COMBINE; no, I don’t think I’ll be able to this year!). (Scroll down that thread for a response by Nicolas Le Novère as to why that isn’t necessarily correct.) So, orthogonality, or the extent to which two or more standards overlap, is sometimes a hard thing to determine.

So, what have I learnt? As always, we must be practical. We should try to develop an elegant solution, but it really, really should be one which is easy to use and intuitive to understand. It’s hard to get to that point, especially as I think that point is (and should be) a moving target. From my perspective, group standards begin with islands of initial research in a field, which then gradually develop into a nascent community. As a field evolves, ‘just-enough’ strategies for storing and structuring data become ‘nowhere-near-enough’. Communication with your peers becomes more and more important, and it becomes imperative that standards are developed.

This may sound obvious, but the practicalities of creating a community standard means such work requires a large amount of effort and continued goodwill. Even with the best of intentions, with every participant working towards the same goal, it can take months – or years – of meetings, document revisions and conference calls to hash out a working standard. This isn’t necessarily a bad thing, though. All voices do need to be heard, and you cannot have a viable standard without input from the community you are creating that standard for. You can have the best structure or semantics in the world, but if it’s been developed without the input of others, you’ll find people strangely reluctant to use it.

Every time I take part in a new standard, I see others like me who have themselves been involved in the creation of standards. It’s refreshing and encouraging. Hopefully the time it takes to create standards will drop as the science community as a whole gets more used to the idea. When I started, the only real standards in biological data (at least that I had heard of) were the structures defined by SWISS-PROT and EMBL/GenBank/DDBJ. By the time I left the EBI in 2006, I could have given you a list a foot long (GO, PSI, and many others), and that list continues to grow. Community engagement and cross-community discussions continue to be popular.

In this context, I can now add synthetic biology standards to my list of standards I’ve been involved in. And, as much as I’ve seen new communities and new standards, I’ve also seen a large overlap in the standardization efforts and an even greater willingness for lots of different researchers to work together, even taking into account the sometimes violent disagreements I’ve witnessed! The more things change, the more they stay the same…

At this stage, it is just a limited involvement, but the BBSRC Synthetic Biology Standards Workshop I’m involved in today and tomorrow is a good place to start with synthetic biology. I describe most of today’s talks in this post, and will continue with another blog post tomorrow. Enjoy!

For those with less time, here is a single sentence for each talk that most resounded with me:

Mike Cooling: Emphasising the ‘re’ in reusable, and make it easier to build and understand large models from reusable components.

Neil Wipat: For a standard to be useful, it must be computationally amenable as well as useful for humans.

Herbert Sauro: Currently there is no formal ontology for synthetic biology, but one will need to be developed.

This meeting is organized by Jen Hallinan and Neil Wipat of Newcastle University. Its purpose is to set up key relationships in the synthetic biology community to aid the development of a standard for that community. Today, I listened to talks by Mike Cooling, Neil Wipat, and Herbert Sauro. I was – unfortunately – unable to be present for the last couple of talks, but will be around again for the second – and final – day of the workshop tomorrow.

Mike Cooling – Bioengineering Institute Auckland, New Zealand

Mike uses CellML (it’s made where he works, but that’s not the only reason…) in his work with systems and synthetic biology models. Among other things, it wraps MathML and partitions the maths, variables and units into reusable pieces. Although many of the parts seem domain specific, CellML itself is actually not domain specific. Further, unlike other modelling languages such as SBML, components in CellML are reusable and can be imported into other models. (Yes, a new package called comp in SBML Level 3 is being created to allow the importing of models into other models, but it isn’t mature – yet.)

They start with template mathematical structures (which are little parts of CellML), and then use the import functionality available as part of CellML to combine the templates into larger physical things/processes (‘SVPs’) and ultimately to combine things into system models.

They extended the CellMLRepository to hold the resulting larger multi-file models, which included adding a method of distributed version control and allow the sharing of models between projects through embedded workspaces.

What can these pieces be used for? Some of this work included the creation of a CellML model of the biology represented in Levskaya et al. 2005 and deposit all of the pieces of the model in the CellML repository. Another example is a model he’s working on about shear stress and multi-scale modelling for aneurysms.

Modules are being used and are growing in number, which is great, but he wants to concentrate more at the moment on the ‘re’ of the reusable goal, and make it easier to build and understand large models from reusable components. Some of the integrated services he’d like to have: search and retrieval, (semi-automated) visualization, semantically-meaningful metadata and annotations, and semi-automated composition.

All this work above converges on the importance of metadata. With the CellML Metadata Framework 1.0, not many used it. With version 2.0 they have developed a core specification with is very simple and then provide many additional satellite specifications. For example, there is a biological information satellite, where you use the biomodels qualifiers as relationships between your data and MIRIAM URNs. The main challenge is to find a database that is at the right level of abstraction (e.g. canonical forms of your concept of interest).

Neil Wipat – Newcastle University

Please note Neil Wipat is my PhD supervisor.

Speaking about data standards, tool interoperability, data integration and synthetic biology, a.k.a “Why we need standards”. They would like to promote interoperability and data exchange between their own tools (important!) as well as other tools. They’d also like to facilitate data integration to inform the design of biological systems both from a manual designer’s perspective and from the POV of what is necessary for computational tool use. They’d also like to enable the iterative exchange of data and experimental protocols in the synthetic biology life cycle.

A description of some of the tools developed in Neil’s group (and elsewhere) exemplify the differences in data structures present within synthetic biology. BacilloBricks was created to help get, filter and understand the information from the MIT registry of standard parts. They also created the Repository of Standard Virtual Biological Parts. This SVP repository was then extended with parts from Bacillus and was extended to make use of SBML as well as CellML. This project is called BacilloBricks Virtual. All of these tools use different formats.

It’s great having a database of SVPs, but you need a way of accessing and utilizing the database. Hallinan and Wipat have started a collaboration with Microsoft Research with the people who created a programming language for genetic engineering of living cells called the genetic engineering of cells (GEC) simulator. Some work a summer student did created a GEC compiler for SVPs from BacilloBricks virtual. Goksel has also created the MoSeC system where you can automatically go from a model to a graph to a EMBL file.

They also have BacillusRegNet, which is an information repository about transcription factors for Bacillus spp. It is also a source of orthogonal transcription factors for use in B. subtilis and Geobacillus. Again, it is very important to allow these tools to communicate efficiently.

The data warehouse they’re using is ONDEX. They feed information from the ONDEX data store to the biological parts database. ONDEX was created for systems biology to combine large experimental datasets. ONDEX views everything as a network, and is therefore a graph-based data warehouse. ONDEX has a “mini-ontology” to describe the nodes and edges within it, which makes querying the data (and understanding how the data is structured) much easier. However, it doesn’t include any information about the synthetic biology side of things. Ultimately, they’d like an integrated knowledgebase using ONDEX to provide information about biological virtual parts. Therefore they need a rich data model for synthetic biology data integration (perhaps including an RDF triplestore).

Interoperabiligy, Design and Automation: why we need standards.

Requirement 1. There needs to be interoperability and data exchange among these tools as well as among these tools and other external tools. Requirement 2. Standards for data integration aid the design of synthetic systems. The format must be both computationally amenable and useful for humans. Requirement 3. Automation of the design and characterization of synthetic systems, and this also requires standards.

The requirements of synthetic biology research labs such as Neil Wipat’s make it clear that standards are needed.

KEYNOTE: Herbert Sauro – University of Washington, US

Herbert Sauro described the developing community within synthetic biology, the work on standards that has already begun, and the Synthetic Biology Open Language (SBOL).

He asks us to remember that Synthetic Biology is not biology – it’s engineering! Beware of sending synthetic biology grant proposals to a biology panel! It is a workflow of design-build-test. He’s mainly interested in the bit between building and testing, where verification and debugging happens.

What’s so important about standards? It’s critical in engineering, where if increases productivity and lowers costs. In order to identify the requirement you must describe a need. There is one immediate need: store everything you need to reconstruct an experiment within a paper (for more on this see the Nature Biotech paper by Peccoud et al. 2011: Essential information for synthetic DNA sequences). Currently, it’s almost impossible to reconstruct a synthetic biology experiment from a paper.

There are many areas requiring standards to support the synthetic biology workflow: assembly, design, distributed repositories, laboratory parts management, and simulation/analysis. From a practical POV, the standards effort needs to allow researchers to electronically exchange designs with round tripping, and much more.

The standardization effort for synthetic biology began with a grant from Microsoft in 2008 and the first meeting was in Seattle. The first draft proposal was called PoBoL but was renamed to SBOL. It is a largely unfunded project. In this way, it is very similar to other standardization projects such as OBI.

DARPA mandated 2 weeks ago that all projects funded from Living Foundries must use SBOL.

SBOL is involved in the specification, design and build part of the synthetic biology life cycle (but not in the analysis stage). There are a lot of tools and information resources in the community where communication is desperately needed.

SBOL Semantic, SBOL Visual, and SBOL Script. SBOL Semantic is the one that’s going to be doing all of the exchange between people and tools. SBOL Visual is a controlled vocabulary and symbols for sequence features.

Have you been able to learn anything from SBML/SBGN, as you have a foot in both worlds? SBGN doesn’t address any of the genetic side, and is pretty complicated. You ideally want a very minimalistic design. SBOL semantic is written in UML and is relatively small, though has taken three years to get to this point. But you need host context above and beyond what’s modelled in SBOL Semantic. Without it, you cannot recreate the experiment.

Feature types such as operator sites, promoter sites, terminators, restriction sites etc can go into the sequence ontology (SO). The SO people are quite happy to add these things into their ontology.

SBOLr is a web front end for a knowledgebase of standard biological parts that they used for testing (not publicly accessible yet). TinkerCell is a drag and drop CAD tool for design and simulation. There is a lot of semantic information underneath to determine what is/isn’t possible, though there is no formal ontology. However, you can semantically-annotate all parts within TinkerCell, allowing the plugins to interpret a given design. A TinkerCell model can be composed of sub-models. Makes it easy to swap in new bits of models to see what happens.

WikiDust is a TinkerCell plugin written in Python which searches SBPkb for design components, and ultimately uploads them to a wiki. LibSBOLj is a library for developers to help them connect software to SBOL.

The physical and host context must be modelled to make all of this useful. By using semantic web standards, SBOL becomes extensible.

Currently there is no formal ontology for synthetic biology but one will need to be developed.

Please note that the notes/talks section of this post is merely my notes on the presentation. I may have made mistakes: these notes are not guaranteed to be correct. Unless explicitly stated, they represent neither my opinions nor the opinions of my employers. Any errors you can assume to be mine and not the speaker’s. I’m happy to correct any errors you may spot – just let me know!

What’s going on these days in the world of reasoning and systems biology modelling? What were people’s experiences when trying to reason over systems biology data in BioPAX and/or SBML format? These were the questions that Andrea Splendiani wanted to answer, and so he collected three of us with some experience in the field to give 10-minute presentations to interested parties at a BioPAX telecon. About 15 people turned up for the call, and there were some very interesting talks. I’ll leave you to decide for yourselves if you’d class my presentation as interesting: it was my first talk since getting back from leave, and so I may have been a little rusty!

The first talk was given by Michel Dumontier, and covered some recent work that he and colleagues performed on converting SBML to OWL and reasoning over the resulting entities.

Essentially, with the SBMLHarvester project, the entities in the resulting OWL file can be divided into two broad categories: in silico entites covering the model elements themselves, and in vivo entities covering the (generally biological) subjects the model elements represent. They copied all of BioModels into the OWL format and performed reasoning and analysis over the resulting information. Inconsistencies were found in the annotation of some of the models, and additionally queries can be performed over the resulting data set.

I gave the second talk about my experiences a few years ago converting SBML to OWL using Model Format OWL (MFO) (paper) and then, more recently, using MFO as part of a larger semantic data integration project whose ultimate aim is to annotate systems biology models as well as create skeleton (sub)models.

I first started working on MFO in 2007, and started applying that work to the wider data integration methodology called rule-based mediation (RBM) (paper) in 2009. As with SBMLHarvester, libSBML and the OWLAPI are used in the creation of the OWL files based on BioModels entries. All MFO entries can be reasoned over and constraints present within MFO from the SBML XSD, the SBML Manual, and from SBO do provide some useful checks on converted SBML entries. The semantics of SBMLHarvester are more advanced than that of MFO, however MFO is intended to be a conversion of a format only, so that SWRL mappings can be used to input/output data from MFO to/from the core of the rule-based mediation. Slide 8 of the above presentation provides a graphic of how rule-based mediation works. In summary, you start with a core ontology which should be small and tightly-scoped to your biological domain of interest. Data is fed to the core from multiple syntactic ontologies using SWRL mappings. These syntactic ontologies can be either direct format conversions from other, non-OWL, formats or pre-existing ontologies in their own right. I use BioPAX in this integration work, and while I have mainly reasoned over MFO (and therefore SBML), I do also work with BioPAX and plan to work more with it in the near future.

The final presenter was Ismael Navas Delgado, whose presentation is available from Dropbox. His talk covered two topics: reasoning over BioPAX data taken from Reactome, and the use of a database back-end called DBOWL for the OWL data. By simply performing reasoning over a large number of BioPAX entries, Ismael and colleagues were able to discover not just inconsistencies in the data entries themselves, but also in the structure of BioPAX. It was a very interesting summary of their work, and I highly recommend looking over the slides.

And what is the result of this TC? Andrea has suggested that, after discussion on the mailing list (contact Andrea Splendiani if you are not on it and want to be added) and then have another TC in a couple of weeks. Andrea has also suggested that it would be nice to “setup a task force within this group to prepare a proof of concept of reasoning on BioPAX, across BioPAX/SBML, or across information resources (BioPAX/OMIM…)”. I think that would be a lot of fun. Join us if you do too!

In the past week, I’ve loaded all curated BioModels from the June release into MFO: that’s over 84,000 individuals!1 It takes a few minutes, but it is possible to view all of those files in Protege 3.4 or higher. However, I’m still trying to work out the fastest way to reason over all those individuals at once. Pellet 2.0.0 rc7 performs the slowest over MFO, and FaCT++ the fastest. I’ve got a few more reasoners to try out, too. Details of reasoning times can be found in the MFO subverison project.

Why SBML in OWL?

Jupiter and its biggest moons (not shown to scale). Public Domain, NASA.

For my PhD, I’ve been working on a semantic data integration. Imagine a planet and its satellites: the planet is your specific domain of biological interest, and the satellites are the data sources you want to pull information from. Then, replace the planet with a core ontology that richly describes your domain of biology in a semantically-meaningful way. Finally, replace each of those satellite data sources with OWL representations, or syntactic ontologies of the format in which your data sources are available. By layering your ontologies like this, you can separate out the process of syntactic integration (the conversion of satellite data into a single format) from the semantic integration, which is the exciting part. Then you can reason over, query, and browse that core ontology without needing to think about the format all that data was once stored in. It’s all presented in a nice, logical package for you to explore. It’s actually very fun. And slowly, very slowly, it’s all coming together.

Really, why SBML in OWL?

As one of my data sources, I’m using BioModels. This is a database of simulatable, biological models whose primary format is SBML. I’m especially interested in BioModels, as the ultimate point of this research is to aid the modellers where I work in annotating and creating new models. In BioModels, the “native” format for the models is SBML, though other formats are available. Because of the importance of SBML in my work, MFO is one of the most important of my syntactic “satellite” ontologies for rule-based mediation.

How a single reaction looks in MFO when viewed with Protege 3.4.

How a single species looks in MFO when viewed with Protege 3.4.

Is this all MFO is good for?

No, you don’t need to be interested in data integration to get a kick out of SBML in OWL: just download the MFO software package, pick your favorite BioModels curated model from the src/main/resources/owl/curated-sbml/singletons directory, and have a play with the file in Protege or some other OWL editor. All the details to get you started are available from the MFO website. I’d love to hear what you think about it, and if you have any questions or comments.

MFO is an alternative format for viewing (though not yet simulating) SBML models. It provides logical connections between the various parts of a model. It’s purpose is to be a direct translation of SBML, SBO, and the SBML Specification document in OWL format. Using an editor such as Protege, you can manipulate and create models, and then using the MFO code you can export the completed model back to SBML (while the import feature is complete, the export feature is not yet finished, but will be shortly).

For even more uses of MFO, see the next section.

Why not BioPAX?

All BioModels are available in it, and it’s OWL!

BioPAX Level 3, which isn’t broadly used yet, has a large number of quite interesting features. However, I’m not forgetting about BioPAX: it plays a large role in rule-based mediation for model annotation (more on that in another post, perhaps). It is a generic description of biological pathways and can handle many different types of interactions and pathway types. It’s already in OWL. BioModels exports its models in BioPAX as well as SBML. So, why don’t I just use the BioPAX export? There are a few reasons:

Most importantly, MFO is more than just SBML, and the BioPAX export isn’t. As far as I can tell, the BioModels BioPAX export is a direct conversion from the SBML format. This means it should capture all of the information in an SBML model. But MFO does more than that – it stores logical restrictions and axioms that are only otherwise stored in either SBO itself or, more importantly, the purely human-readable content from the SBML specification document2. Therefore MFO is more than SBML, it is a bunch of extra constraints that aren’t present in the BioPAX version of SBML, and therefore, I need MFO as well as BioPAX.

I’m making all this for modellers, especially those who are still building their models. None of the modellers at CISBAN, where I work, natively use BioPAX. The simulators accept SBML. They develop and test their models in SBML. Therefore I need to be able to fully parse and manipulate SBML models to be able to automatically or semi-automatically add new information to those models.

Export of data from my rule-based mediation project needs to be done in SBML. The end result of my PhD work is a procedure that can create or add annotation to models. Therefore I need to export the newly-integrated data back to SBML. I can use MFO for this, but not BioPAX.

For people familiar with SBML, MFO is a much more accessible view of models than BioPAX. If you wish to start understanding OWL and its benefits, using MFO (if you’re already familiar with SBML) is much easier to get your head around.

What about CellML?

You call MFO “Model” Format OWL, yet it only covers SBML.

Yes, there are other model formats out there. However, as you now know, I have special plans for BioPAX. But there’s also CellML. When I started work on MFO more than a year ago, I did have plans to make a CellML equivalent. However, Sarala Wimalaratne has since done some really nice work on that front. I am currently integrating her work on the CellML Ontology Framework. She’s got a CellML/OWL file that does for CellML what MFO does for SBML. This should allow me to access CellML models in the same way as I can access SBML models, pushing data from both sources into my “planet”-level core ontology.

It’s good times in my small “planet” of semantic data integration for model annotation. I’ll keep you all updated.

Footnotes:

1. Thanks to Michael Hucka for adding the announcement of MFO 2 to the front page of the SBML website!.
2. Of course, not all restrictions and rules present in the SBML specification are present in MFO yet. Some are, though. I’m working on it!